Adolescent ; Chondrosarcoma ; pathology ; surgery ; Douglas' Pouch ; pathology ; surgery ; Female ; Humans ; Peritoneal Neoplasms ; pathology ; surgery ; Soft Tissue Neoplasms ; pathology ; surgery

To investigate the secondary metabolites of endophytic fungi Pericinia sp. F-31. Column chromatography on silica gel, Sephadex LH-20 and semi-preparative HPLC were used to separate and purify the compounds. Two compounds were isolated from the fermentation broth of Periconia sp. Their structures were identified as 5-(1-hydroxyhexyl) -6-methyl-2H-pyran-2-one (1) and 2-(3-hydroxy-4-methylphenyl) -propanoic acid (2). Compound 1 was a new lactone compound, compound 2 was new natural product, and the NMR data of compound 2 was reported for the first time.
Annona ; microbiology ; Ascomycota ; chemistry ; genetics ; isolation & purification ; metabolism ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; metabolism ; Endophytes ; chemistry ; genetics ; isolation & purification ; metabolism ; Lactones ; chemistry ; isolation & purification ; metabolism ; Mass Spectrometry ; Molecular Structure

Objective The relationship between circular RNAs (circRNA) and lung adenocarcinoma (LAC) remains to be further explored. This study aimed to investigate the expression of Hsa_circ_0002360 in LAC and its value in the clinical diagnosis and treatment of the malignancy. Methods This study included 50 cases of LAC treated in our hospital from May 2018 to June 2019 and another 50 healthy subjects as normal controls. We determined the expression of Hsa_circ_0002360 in the LAC cells by qRT⁃PCR. We cultured human LAC cell lines A549, H1299 and H1975 and human bronchial mucosal epithelial cell line BEAS-2B, inoculated the A549 and H1975 LAC cells in the 6-well plate, with 3 wells for si-circRNAs and the other 3 for negative controls (si-NC), followed by cell transfection experiment. Using the CCK8, colony formation and wound healing assays, we detected the effects of Hsa_circ_0002360 on the activity, proliferation, invasiveness and metastasis of the A549 and H1975 cells. Results The expression of Hsa_circ_0002360 was significantly higher in the LAC tissue than in the adjacent tissue (2.84 ± 0.12 vs 1.46 ± 0.08, P < 0.01), and so was it in the serum of the LAC patients than in the normal controls (2.59 ± 0.17 vs 1.21 ± 0.11, P < 0.01), but remarkably lower in the serum of the patients after than before surgery (1.58 ± 0.09 vs 2.65 ± 0.13, P < 0.01). Besides, its expression was significantly increased in the LAC A549, H1299 and H1975 cell lines compared with that in the BEAS-2B cells (P < 0.05). In comparison with the si-NC group, the si-circRNA group showed significant decreases in the rates of colony formation ([65.66± 4.93]% vs [38.33 ± 1.69]%, P < 0.01) and migration of the cells ([54.66±4.10]% vs [13.50 ± 3.34]%, P < 0.01) and in the count of H1975 cells as well (P < 0.01). Conclusion Hsa_circ_0002360 can be used as a biomarker for the early diagnosis and treatment of lung adenocarcinoma.

In the research and development of new drugs, it is very important to investigate the in vitro metabolism of candidate drugs. Traditional models such as liver microsomes have many limitations, while the in vitro model of recombinant human drug metabolizing enzymes is considered as an important and useful approach because of its convenient access, stable activity and low cost. In this study, six major human UDP-glucuronosyltransferases (UGTs) genes (UGT1A1, 1A3, 1A4, 1A6, 1A9 and 2B7) were cloned from human liver cDNA and heterologously expressed in Saccharomyces cerevisiae and baculovirus-infected insect cell. UGT1A1, 1A3, 1A6 and 1A9 were successfully expressed in yeast and showed glucuronidation activity against a variety of different structural types of substrates, but their activities were low. All six UGTs were successfully expressed and exhibited significantly improved glucuronidation activity when Trichopolusia ni cells BTI-TN5B1-4 (High Five) were used as the host. The recombinant human UGTs expressed in insect cells can catalyze the glucuronidation of their specific substrates, and the glucuronidation products were synthesized at milligram-scale with yields of 13%-66% for the first time, of which the structures were identified via MS, ¹H NMR, and ¹³C NMR spectroscopic analysis. Above all, the recombinant human UGTs yeast and insect cell expression systems constructed in this study can be used for in vitro metabolism evaluation in the early stage of new drugs research and development, and also provide a new tool for the synthesis of glucuronide metabolites.

OBJECTIVETo investigate the protein expression of caveolin-1 in type II alveolar epithelial cells (A549) exposed to carbon black nanoparticles (CB NPs) and the role of caveolin in the endocytosis of CB NPs.

METHODSA549 cells were exposed to 0, 25, 50, 100, 200, and 400 µg/ml CB NPs for 24 h; then, trypan blue assay was applied to determine the cell viability. A549 cells were also exposed to 0, 25, 50, and 100 µg/ml CB NPs for 24 h, then, transmission electron microscopy (TEM) and flow cytometry were applied to observe the morphological change of cells and cellular side scatter (SSC), and Western blot was used to analyze the effect of CB NPs on the protein expression of caveolin-1. A549 cells were co-exposed to1 µg/ml filipin and 100 µg/ml CB NPs for 24 h, then, the cellular SSC was observed.

RESULTSCompared with controls, the A549 cells exposed to 200 and 400 µg/ml CB NPs had the cell viability decreased by 38.2% and 46.6%, respectively (P < 0.05), while those exposed to 25, 50, and 100 µg/ml CB NPs showed no significant decrease in cell vitality (P > 0.05). The protein expression of caveolin-1 was significantly higher in the cells exposed to 50 and 100 µg/ml CB NPs than in controls (P < 0.05). The TEM showed that plasmalemmal vesicles containing black particles were found in the cytoplasm of the cells exposed to 50 and 100 µg/ml CB NPs. The flow cytometry showed that the cellular SSC ratio increased from 1.007 to 1.331 as the dose of CB NPs rose within 0 ∼ 100 µg/ml and fell to 1.25 after the cells were co-exposed to1 µg/ml filipin and 100 µg/ml CB NPs.

CONCLUSIONCarbon black nanoparticles can be transferred into A549 cells by endocytosis, but caveolin-mediated endocytic pathway plays a minor role in this process.

Caveolin 1 ; physiology ; Cell Line ; Endocytosis ; Humans ; Nanoparticles ; Soot ; pharmacokinetics

OBJECTIVETo investigate the biochemical metabolic changes detected by phosphorus-31 MR spectroscopy ((31)P MRS) with pathologic changes in the liver of fasting rabbits.

METHODSA total of 22 rabbits were under the starvation up to death to establish animal models. Hepatic (31)P MRS was performed in different period of 10 rabbits including normal condition, over-starvation, agonal condition and death after 30 min. Other 9 rabbits were divided into three type including over-starvation, agonal condition and death group with 3 rabbits in each group, and 3 healthy rabbits served as controls. All the 12 rabbits were sacrificed for the hepatic pathological examination. The MR examination was performed on a 1.5 T imager using a 1H/31P surface coil by the 2D chemical shift imaging technique. The relative quantities of phosphomonoesters (PME), phosphodiesters (PDE), inorganic phosphate (Pi) and adenosine triphosphate (ATP) were measured.

RESULTSAll the relative quantification of phosphorus metabolites were changed significantly from starvation to death (X(2)=23.13-35.41, P<0.01). The relative quantifications of ATP of normal condition, over-starvation, agonal condition and death were 2.54 +/-0.53, 1.73 +/-0.14, 0.88 +/-0.23 and 0.05 +/-0.08, respectively (rs=1.0, P<0.01). The relative quantifications of PDE from normal to death were 1.25 +/-0.54, 2.76 +/-0.23, 3.33 +/-0.49 and 3.87 +/-0.43, respectively, and those of Pi were 0.42 +/-0.02, 0.65 +/-0.05, 0.89 +/-0.15 and 0.99 +/-0.08, respectively (rs=1.0, P <0.01). The relative quantifications of PME were also significantly changed (rs=0.4, P=0.6). The pathologic changes of normal condition, over-starvation, agonal condition and death: decreased size of hepatocytes, loss of cell number, cellular swelling, degeneration and cell necrosis or hepatic hemorrhage became more and more pronounced.

CONCLUSION(31)P MRS can monitor dynamic changes of relative quantification of phosphorus metabolites, which are correlated with the pathological severity of acute hepatic injury by fasting.

Animals ; Death ; Dose-Response Relationship, Radiation ; Female ; Liver ; metabolism ; pathology ; Magnetic Resonance Spectroscopy ; methods ; Male ; Phosphorus ; metabolism ; Phosphorus Isotopes ; metabolism ; Rabbits ; Random Allocation ; Starvation

OBJECTIVE: To investigate the effect of diet-induced hyperlipidemia on TGF-beta/Smad signaling pathway in the kidney of diabetic rats, and to explore the mechanism by which hyperlipidemia leads to renal injury in diabetes. METHODS: Diabetic rats and non-diabetic rats were fed with normal fat diet and high fat diet for 16 weeks, respectively. The expressions of TGF-beta1, TbetaRII, and Col-IV mRNA in the renal cortex were examined by reverse transcriptase-PCR,TbetaRII and p-Smad staining in glomerular cells were detected by immunohistochemical staining, and the expression of TGF-beta1 and Col-IV protein was determined by Western blot. RESULTS: Diet-induced hyperlipidemia up-regulated the levels of TGF-beta1, TbetaRII, p-Smad, and Col-IV protein and mRNA in the renal cortex of diabetic rats compared with those of non-diabetic rats. However, feeding high fat diet to non-diabetic rats had no influence on the expression of TGF-beta1, TbetaRII, p-Smad2, and Col-IV in the renal cortex. CONCLUSION Hyperlipidemia induced by high fat diet ingestion leads to renal injury in diabetic rats through activating TGF-beta1 /Smad signaling pathway.
Animals ; Diabetes Mellitus, Experimental ; complications ; metabolism ; Diabetic Nephropathies ; metabolism ; Dietary Fats ; administration & dosage ; Female ; Hyperlipidemias ; complications ; metabolism ; Kidney ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Smad Proteins ; metabolism ; Transforming Growth Factor beta1 ; metabolism

Based on the principle of the pathway engineering, a novel pathway of producing glycerol was built in E. coli. The gpd1 gene encoding glycerol 3-phosphate dehydrogenase and the hor2 gene encoding glycerol 3-phosphatase were cloned from Saccharomyces cerevisiae, respectively. The two genes were inserted into expression vector pSE380 together. A recombinant plasmid pSE-gpd1-hor2 containing polycistron was constructed under the control of the strong trc promoter. Then it was transformed into E. coli BL21. The result showed the recombinant microorganism GxB-gh could convert glucose to glycerol directly. And the recombinant microorganism GxB-gh was incubated to produce glycerol from D-glucose in the fermentor. The maximal concentration of glycerol was 46.67g/L at 26h. Conversion rate of glucose was 42.87%. The study is about "green" producing glycerol by recombinant microorganism and is also useful for further working in recombining microorganism of producing 1,3-propanediol.
Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Fermentation ; Fungal Proteins ; biosynthesis ; genetics ; Genetic Engineering ; Glycerol ; metabolism ; Glycerolphosphate Dehydrogenase ; biosynthesis ; genetics ; Phosphoric Monoester Hydrolases ; biosynthesis ; genetics ; Saccharomyces cerevisiae ; enzymology ; genetics

OBJECTIVEThis study was conducted to study the effects of sodium selenite on rat hepatocellular DNA damage, apoptosis, changes of cell cycle and DNA relative content induced by cadmium chloride in vivo.

METHODSBoth sodium selenite at the dose of 5 micromol/kg and cadmium chloride at the dose of 5 micromol/kg, 10 micromol/kg and 20 micromol/kg were given to rats by i.p. and there were 5 male SD rats in each group. Hepatocellular DNA damage was measured by the single cell gel electrophoresis (or comet assay), hepatocellular apoptosis was measured with TUNEL (TdT-mediated dUTP Nick End Labelling) and flow cytometry, DNA relative content (DNA(RC)) and cell cycle were detected with flow cytometry.

RESULTSWhen sodium selenite at the dose of 5 micromol/kg acted jointly with cadmium chloride at the dose of 5 micromol/kg, 10 micromol/kg and 20 micromol/kg respectively, the results showed that selenium reduced the effect of cadmium on DNA damage and apoptosis and decreased the rates of DNA damage and the rates of apoptosis significantly. Sodium selenite at the dose of 5 micromol/kg increased cell number of G(0)/G(1) period decreased by cadmium chloride at the dose of 5 micromol/kg and increased cell number of G(2)/M period decreased by cadmium chloride at the dose of 10 micromol/kg and 20 micromol/kg. Sodium selenite at the dose of 5 micromol/kg increased DNA relative content reduced by cadmium chloride at the dose of 10 micromol/kg and 20 micromol/kg.

CONCLUSIONSIt was suggested that selenium at certain doses could antagonize DNA damage, apoptosis, changes of cell cycle and DNA relative content induced by cadmium in rat hepatocytes in vivo.

Animals ; Apoptosis ; drug effects ; Cadmium Chloride ; antagonists & inhibitors ; toxicity ; Cell Cycle ; drug effects ; Cell Division ; drug effects ; DNA Damage ; drug effects ; Dose-Response Relationship, Drug ; Hepatocytes ; drug effects ; metabolism ; pathology ; Male ; Rats ; Rats, Sprague-Dawley ; Sodium Selenite ; pharmacology

OBJECTIVESThis study was conducted to explore effects of selenium on rat hepatocellular DNA damage induced by cadmium in vitro.

METHODSodium selenite was added at concentrations of 8.75, 17.50 and 35.00 micromol/L respectively with cadmium chloride at the concentrations of 8.75, 17.50 and 35.00 micromol/L respectively and rat hepatocellular DNA damage was measured with single cell gel electrophoresis (comet assay).

RESULTSSodium selenite at the concentration of 8.75 micromol/L inhibited DNA damage caused by cadmium chloride at the concentration of 8.75, 17.50 and 35.00 micromol/L in rat liver cells (P < 0.05). Although sodium selenite at 17.50 micromol/L inhibited DNA damage induced by cadmium chloride at 17.50 and 35.00 micromol/L, it did not inhibit DNA damage induced by cadmium chloride at 8.75 micromol/L. Sodium selenite at 35.00 micromol/L did not have antagonistic effects on DNA damage induced by cadmium chloride at 8.75, 17.50 and 35.00 micromol/L. In addition, sodium selenite at 8.75 micromol/L had the best antagonistic effect while cadmium chloride at 8.75 micromol/L, but the antagonistic effect of sodium selenite at 17.50 micromol/L was better than 8.75 micromol/L while cadmium chloride at 17.50 and 35.00 micromol/L.

CONCLUSIONThe antagonistic effect of selenium on rat hepatocellular DNA damage induced by cadmium related to the concentrations of selenium and also to the concentration ratio between selenium and cadmium.

Animals ; Cadmium ; toxicity ; Comet Assay ; DNA ; drug effects ; genetics ; DNA Damage ; drug effects ; Dose-Response Relationship, Drug ; Hepatocytes ; cytology ; drug effects ; metabolism ; Rats ; Selenium ; pharmacology